Fig. 4.

miR-324-5p was a target of TPT1-AS1 in CC. a Bioinformatics analysis showed that miR-324-5p could directly target 3′-UTR of TPT1-AS1-wild type (WT). TPT1-AS1-mutant (Mut) means mutation of binding sites in the 3′-UTR of TPT1-AS1. b Real-time PCR showed that TPT1-AS1 could negatively regulate miR-324-5p expression in CC cells. c Pearson correlation analysis revealed that an obvious negative association between miR-324-5p and TPT1-AS1 expression in CC tissues. d Dual luciferase reporter assays showed that miR-324-5p could negatively regulate the luciferase activity of TPT1-AS1-WT, rather than TPT1-AS1-Mut. e The association between TPT1-AS1, miR-324-5p and Ago2 was ascertained by analyzing C33A cell lysates using RNA immunoprecipitation with an Ago2 antibody. Real-time PCR was used to detect the TPT1-AS1 level change in the substrate of RIP assay in miR-324-5p-overexpressing CC cells. f Detection of TPT1-AS1 using real-time PCR in the sample pulled down by biotinylated TPT1-AS1 and negative control (NC) probe. Detection of miR-324-5p using real-time PCR in the same sample pulled down by biotinylated TPT1-AS1 and NC probe. *P < 0.05, **P < 0.01